Association Analysis Between Methylation of SCARB1 Gene Promoter and Coronary Heart Disease / 中国医学科学院学报

Objective To explore the relationship between scavenger receptor class B member 1 (SCARB1) gene promoter methylation and the pathogenesis of coronary artery disease. Methods A total of 120 patients with coronary heart disease treated in Renji Hospital affiliated to Shanghai Jiao Tong University School of Medicine from December 2018 to May 2020 were selected as the case group,while 140 gender and age matched healthy participants were randomly selected as the control group for a case-control study.The methylation status was detected by high-throughput target sequencing after bisulfite converting,and the methylation of CpG sites in the promoter region of SCARB1 gene was compared between the two groups. Results The case group showed higher methylation level of SCARB1+67 and lower methylation level of SCARB1+134 than the control group (both P<0.001),and the differences remained statistically significant in men (both P<0.001) and women (both P<0.001).The overall methylation level in the case group was lower than that in the control group [(80.27±2.14)% vs.(81.11±1.27)%;P=0.006],while this trend was statistically significant only in men (P=0.002). Conclusion The methylation of SCARB1 gene promotor is associated with the pathogenesis and may participate in the occurrence and development of coronary heart disease.

Effect of Shuangyu Tiaozhi Decoction on SRB1/CYP7A1/FXR Signaling Pathway in Liver of Hypercholesterolemic Rats / 中国实验方剂学杂志

Objective:To observe the effect of Shuangyu Tiaozhi decoction on B-type scavenger receptor （SRB1）/cholesterol 7<italic>α</italic>-hydroxylase protein （CYP7A1）/farnesol X receptor （FXR） signaling pathway in liver of hypercholesterolemic rats， and its mechanism in reducing blood lipid. Method:Among 40 SD rats， 8 were randomly selected as normal group， and the remaining 32 were successfully established as hypercholesterolemic model， and randomly divided into 4 groups： model group， low and high-dose Shuangyu Tiaozhi decoction groups （7.8， 15.6 g·kg^-1）， and simvastatin group （4 mg·kg^-1）， with 8 rats in each group. The drugs were continuously given for 8 weeks. Serum total cholesterol （TC）， triglyceride （TG） and liver TC，free cholesterol （FC） and total bile acid （TBA） were measured. The pathomorphological changes in liver were observed by Hematoxylin and eosin （HE） Staining. The mRNA and protein expressions of SRB1， CYP7A1 and FXR were determined by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot. The immunohistochemistry was used to detect CYP7A1 and FXR expressions in liver. Result:Compared with the normal group， TC， TG， FC levels in the model group were significantly increased， while the TBA level was markedly decreased， the morphology showed obvious liver steatosis， and significant declines in expressions of SRB1， CYP7A1， FXR were observed by Real-time PCR， Western blot and immunohistochemistry assays （<italic>P</italic><0.05， <italic>P</italic><0.01）. Compared with the model group， the levels of TC，TG，FC in each treatment group were reduced significantly， and the TBA level was increased markedly， the liver steatosis decreased significantly， the results of Real-time PCR， Western blot and immunohistochemistry assays showed significant increase in the expressions of SRB1， CYP7A1， FXR （<italic>P</italic><0.05， <italic>P</italic><0.01）. The therapeutic effect of high-dose Shuangyu Tiaozhi decoction group was more remarkable than that in low-dose Shuangyu Tiaozhi Decoction group （<italic>P</italic><0.05）， with no obvious difference compared with simvastatin group. Conclusion:Shuangyu Tiaozhi decoction can promote hepatic RCT and synthesize bile acid by up-regulating SRB1/CYP7A1/FXR signaling pathway， so as to reduce the blood lipid levels and improve hepatic lipid metabolism of hypercholesterolemic rats.

Arsenic on cholesterol efflux and the expression of ABCA1, ABCG1 and SRBI / 预防医学

Objective@#To explore the impact of arsenic on cholesterol efflux and the expression of ATP-binding cassette, sub-family A, member 1 ( ABCA1 ), ATP-binding cassette transporter G1 ( ABCG1 ), and scavenger receptor class B member I ( SRBI ) in macrophages, so as to provide the evidence for the mechanism of arsenic induced atherosclerosis.@*Methods@#The human myeloid leukemia mononuclear cells ( THP-1 ), induced by phorbol myristate acetate, and mouse primary macrophages were treated with 0, 0.625, 1.25, 2.5 and 5 μmol/L NaAsO2 for 48 hours. Then the cells treated with 2.5 μmol/L NaAsO2 were changed to arsenic free mediums for 48 hours and collected every 12 hours to analyze the time effect of arsenic. The expression levels of ABCA1, ABCG1 and SRBI were determined by quantitative polymerase chain reaction and western blotting. Cholesterol efflux rates were measured by 3H isotope tracer. @*Results@#Arsenic significantly down-regulated the expression levels of ABCA1 and ABCG1, and cholesterol efflux in a dose-dependent manner. The levels of ABCA1 mRNA decreased by 69% and 72%, the levels of ABCG1 mRNA decreased by 42% and 34%, and the rate of cholesterol efflux decreased by 55% and 59% in THP-1 and mouse primary macrophages cells treated with 5 μmol/L NaAsO2 ( all P<0.05 ). Arsenic had no significant effect on SRBI expression ( all P>0.05 ). Arsenic inhibited ABCA1 expression and cholesterol efflux in THP-1 in a time-dependent manner. Compared with cells before the exposure of arsenic, the level of ABCA1 mRNA and the rate of cholesterol efflux in THP-1 bottomed at 48 hours by 43% and 42%, and gradually recovered when arsenic was removed. @*Conclusions@#Arsenic inhibits cholesterol efflux by down-regulating the expression of ABCA1 and ABCG1 in macrophages.

Scavenger Receptor Class A to E Involved in Various Cancers / 전남의대학술지

Scavenger receptors typically bind to multiple ligands on a cell surface, including endogenous and modified host-derived molecules and microbial pathogens. They promote the elimination of degraded or harmful substances such as non-self or altered-self targets through endocytosis, phagocytosis, and adhesion. Currently, scavenger receptors are subdivided into eight classes based on several variations in their sequences due to alternative splicing. Since recent studies indicate targeting scavenger receptors has been involved in cancer prognosis and carcinogenesis, we will focus on the current knowledge about the emerging role of scavenger receptor classes A to E in cancer progression.

CD36 (SR-B2) as a Target to Treat Lipid Overload-Induced Cardiac Dysfunction

The heart faces the challenge of adjusting the rate of fatty acid uptake to match myocardial demand for energy provision at any given moment, avoiding both too low uptake rates, which could elicit an energy deficit, and too high uptake rates, which pose the risk of excess lipid accumulation and lipotoxicity. The transmembrane glycoprotein cluster of differentiation 36 (CD36), a scavenger receptor (B2), serves many functions in lipid metabolism and signaling. In the heart, CD36 is the main sarcolemmal lipid transporter involved in the rate-limiting kinetic step in cardiac lipid utilization. The cellular fatty acid uptake rate is determined by the presence of CD36 at the cell surface, which is regulated by subcellular vesicular recycling from endosomes to the sarcolemma. CD36 has been implicated in dysregulated fatty acid and lipid metabolism in pathophysiological conditions, particularly high-fat diet-induced insulin resistance and diabetic cardiomyopathy. Thus, in conditions of chronic lipid overload, high levels of CD36 are moved to the sarcolemma, setting the heart on a route towards increased lipid uptake, excessive lipid accumulation, insulin resistance, and eventually contractile dysfunction. Insight into the subcellular trafficking machinery of CD36 will provide novel targets to treat the lipid-overloaded heart. A screen for CD36-dedicated trafficking proteins found that vacuolar-type H⁺-ATPase and specific vesicle-associated membrane proteins, among others, were uniquely involved in CD36 recycling. Preliminary data suggest that these proteins may offer clues on how to manipulate myocardial lipid uptake, and thus could be promising targets for metabolic intervention therapy to treat the failing heart.

Zinc Ion Inhibits Macrophage Foaming during Atherosclerosis / 中山大学学报(医学科学版)

@#【Objective】To investigate the effects of zinc on the formation of atherosclerotic macrophage foaming and plaque formation and its mechanism.【Methods】The macrophage foaming model was established by stimulating THP-1cells with oxLDL. The degree of foaming in different zinc concentrations of 0，30 and 60 μmol/Lwas detected by oil red Ostaining and the intake of lipid by foam cells was measured by DiI-oxLDL fluorescence. The relevant scavenger protein ex⁃pression of CD36，SR-A was detected by immunoblotting. The relative expression level of zinc ion transporters was detect⁃ed by real-time fluorescent quantitative PCR. ApoE-/- mice were randomly divided into 4 groups，the normal feed group（Chow group），the high-fat zinc-deficient group（HFD-ZnD），and the high-fat normal zinc group（HFD），high-fatand zinc-supplement group（HFD-ZnS），blood lipids and the protein of the mice aorta were detected in the 13 week.【Results】Compared with the normal zinc group，the oil red O density increased（P < 0.05），and add zinc ion decreased the intake of the DiI-oxLDL by foam cells（P < 0.01）. In the 0 μmol/L zinc group，the SR-A and CD36 protein expressionin the foam cells increased（P < 0.05）and 15μmol/L Zn2+ treatment before stimulating with oxLDL reduced the contentsof SR-A and CD36 proteins（P < 0.05）. Compared the oxLDL-treated group with the control group，the mRNA expres⁃sion levels of ZIP10，ZIP12 and ZIP14 increased，and the mRNA expression levels of ZIP4，ZIP7 and ZIP8 decreased（P < 0.05）；while the mRNA expression of ZnT4 was up-regulated（P < 0.01），and the mRNA expression of ZnT1 was down-regulated（P < 0.05）. Compared with Chow group，low density lipoprotein cholesterol（LDL-C），total cholesterol（TC）and triglyceride（TG）were increased in HFD group and HFD-ZnD group，respectively（P < 0.05）；HFD-ZnD group High-density lipoprotein cholesterol（HDL-C）was significantly elevated. Moreover，the LDL-C of the HFD-ZnS group was significantly lower than that of the HFD-ZnD group（P < 0.05）. The SR-A protein of the mice aorta of the HFD and HFD-ZnD group increased compared to the Chow group（P < 0.01），HFD-ZnS could restrain the increase（P < 0.05）. Compared with the Chow group，the ratio of plaque area in the aorta to the total arterial lumen area was significantly in⁃creased in the HFD-ZnD group（P < 0.01），and HFD-ZnS significantly inhibited this increase（P < 0.01）.【Conclusions】 Extracellular zinc deficiency aggravates lipid deposition in macrophages，and the mechanism may be regulated by up-reg⁃ulating the scavenger receptor CD36 and SR-A. Zinc ion transporters are involved in macrophage foaming and formation ofarterial plaques. Zinc deficiency can increase LDL-C and promote the increase of arterial plaque induced by high-fat diet.

The expression of oxidized low density lipoprotein and scavenger receptor that binds phospha-tidylserine and oxidized lipoprotein in patients with rheumatoid arthritis / 中华风湿病学杂志

Objective To investigate the expression level of oxidized low density lipoprotein (ox-LDL) and its scavenger receptor scavenger receptor that binds phosphatidylserine and oxidized lipoprotein (SRPSOX) in patients with rheumatoid arthritis (RA),and to explore the relationship between ox-LDL and disease activity.Methods The serum ox-LDL in RA patients and healthy control group were detected by enzymelinked immunosorbent assay (ELISA),as well as the fluidox-LDL in RA,osteoarthritis (OA) and inflammatory arthritis (IA).The expression of SR-PSOX in mixed cells of RA and IApatients was detected by western blot.The expression of serum ox-LDL between RA groupand the control group was analyzed by t-test and non-parametric test.The correlation of serum ox-LDL expression levels in RA patients with C-reactive protein (CRP),erythrocyte sedimentation rate (ESR) and other inflammatory factors and disease activitywas analyzed by Pearson linear regression.Results The expression of ox-LDL in the serum of RA patients was significantly higher than that of normal control group [(3 076±131) mU/ml,(2 334±84) mU/ml,t=4.242,P＜0.01].The expression of ox-LDL in synovial fluid of RA patients was significantly higher than that of the OA group [(4 963±354) mU/ml],(3 956±347) mU/ml,t=2.372,P＜0.05).The expression of SR-PSOX in synovial fluid mixed cells of RA patients was higher than that of the IA group [(4.92±0.18) vs (0.24±0.04),t=33.53,P＜0.01].The expression of ox-LDL in serum of RA patients was negatively correlated with ESR,CRP and overall disease activity DAS28 (r=-9.42,P=0.009;r=-0.35,P=0.029 7;r=0.42,P=0.008 4).The expression of ox-LDL in the serum of RA patients with moderate disease activity was significantly higher than those patients with high disease activity [(3 302±138) mU/ml vs (2 464±228) mU/ml,t=3.335,P＜0.01],however,those with low disease activity and disease remission had higher serum ox-LDL expression but without statistical significant differences.After treated with anti-rheumatic drugs (DMARDs),serum ox-LDL of RA patients had a trend of slight increasing butwithout sign-ificant difference.The ox-LDL/LDL-C or ox-LDL/HDL-C was negatively not correlated with disease activity score in 28 (DAS28),ESR,CRP.Conclusion In RA patients,the expression of ox-LDL in the serum and synovial fluid is high and the SR-PSOX expressionin synovial fluid is also high.The serum ox-LDL levels are negatively correlated with ESR,CRP and DAS28,which are related to disease activity of RA.These findings suggest that the ox-LDL and the receptor SR-PSOX may play a role in RA pathogenesis,but needs further study.

Inhibitory mechanism of timosaponin B-Ⅱagainst proliferation and migration of human gastric cancer cells / 第二军医大学学报

Objective To explore the inhibitory mechanism of timosaponin B-Ⅱ against the proliferation and migration of human gastric cancer cell lines BGC-823 and MGC-803. Methods BGC-823 and MGC-803 cells were treated with timosaponin B-Ⅱ (50 ng/mL) for 48 h, and the mRNA and protein expressions of scavenger receptor A5 (SCARA5) were measured by qPCR and Western blotting, respectively. The binding site of hsa-miRNA-766-3p in SCARA5 gene 3′UTR was predicted by bioinformatics, and was validated by luciferase report assay. After transfecting with hsa-miRNA-766-3p mimic or siRNA-SCARA5 for 24 h, the BGC-823 and MGC-803 cells were treated with timosaponin B-Ⅱ(50 ng/mL) for 48 h. The relative levels of hsamiRNA-766-3p and SCARA5 protein expression were detected by qPCR and Western blotting, respectively. The proliferation and migration abilities of cells were determined by MTT. Results The expressions of SCARA5 protein in BGC-823 and MGC-803 cells treated with timosaponin B-Ⅱ (50 ng/mL) for 48 h were significantly increased versus the control group (P0.01), while no significant difference was found in relative mRNA level of SCARA5 between the timosaponin B-Ⅱ treated cell group and the control group (P0.05). Compared with transfection of reporter gene expression vector alone group, the luciferase activity was significantly inhibited or enhanced in the cells co-transfected with hsa-miRNA-766-3p mimic or hsa-miRNA-766-3p inhibitor and wild-type luciferase reporter gene (P0.05, P0.01). No change was observed between the cells co-transfected with hsa-miRNA-766-3p mimic or hsa-miRNA-766-3p inhibitor and mutant-type luciferase reporter gene expression vector and the cells transfected with reporter gene expression vector alone (P0.05). Hsa-miRNA-766-3p levels were significantly decreased and SCARA5 protein expressions were significantly increased in BGC-823 and MGC-803 cells treated with 50 ng/mL timosaponin B-Ⅱ (P0.01). Compared with the timosaponin B-Ⅱ treatment group, hsa-miRNA-766-3p levels were significantly increased and SCARA5 protein expressions were significantly decreased in the BGC-823 and MGC-803 cells of the hsa-miRNA-766-3p mimic transfection＋ timosaponin B-Ⅱ treatment group (P0.01). There were no differences in the hsa-miRNA-766-3p levels between the hsa-miRNA-766-3p mimic transfection＋timosaponin B-Ⅱtreatment group and the timosaponin B-Ⅱ treatment group (P0.05), but SCARA5 protein expressions were significantly decreased (P0.01). Compared with cell control group and vehicle control group, the proliferation and migration abilities of BGC-823 and MGC-803 cells were significantly inhibited by timosaponin B-Ⅱ (P0.01). Compared with the timosaponin B-Ⅱ treatment group, the proliferation and migration abilities of BGC-823 and MGC-803 cells were significantly increased in the siRNA-SCARA5 transfection＋timosaponin B-Ⅱ treatment group (P0.01). Conclusion Timosaponin B-Ⅱ can inhibit the proliferation and migration of BGC-823 and MGC-803 cells via suppressing hsamiRNA-766-3p and upregulating the target gene SCARA5.

Protective effects of high-density lipoprotein on mice cardiomyocytes induced by oxygen and glucose deprivation through Akt signaling pathway / 中华危重病急救医学

Objective To investigate the protective effect of high-density lipoprotein (HDL) on the mice cardiac myocytes induced by oxygen and glucose deprivation (OGD).Methods Cardiac cells of primary scavenger receptor-B1 knockout mice (SR-B1-/-) and normal C57 mice (SR-B1+/+) were obtained by protease digestion and differential adhesion method. ① The two kinds of cells were divided into normal control group (Con group), OGD group, OGD+HDL group. Propidium iodide (PI) staining were used to determine the necrosis of cardiac myocytes. ② SR-B1+/+cardiac cells were divided into Con group, OGD group, OGD+HDL group, and phosphatidylinositol 3 kinase/protein kinase B (PI3K/Akt) inhibitor LY294002 group. PI staining were used to determine the necrosis of cardiac myocytes. TUNEL staining was used to determine the cell apoptosis. The kit was used to determine the contents of MB isoenzyme of creatine kinase (CK-MB) and lactate dehydrogenase (LDH) in the culture medium supernatant. The expressions of SR-B1 and Akt protein were determined by Western Blot.Results ① In SR-B1+/+ cardiomyocytes, HDL could inhibit cell necrosis induced by OGD. There was no protective effect of HDL on OGD in the SR-B1-/- cardiomyocytes.② The study of SR-B1+/+ cells was showed that compared with Con group, necrotic cells were significantly increased and cell activity were significantly decreased, the cell viability were significantly decreased, the contents of LDH and CK-MB in supernatant were significantly increased, the expressions of phosphorylated Akt (p-Akt) and SR-B1 were significantly decreased in OGD group. Compared with OGD group, the number of necrotic cells in the OGD+HDL group was significantly decreased [PI positive cells rate: (26.71±5.94)％ vs. (64.24±18.34)％], the cell activity was significantly increased [(63.84±6.95)％ vs. (26.71±5.13)％], the contents of LDH and CK-MB in supernatant were significantly decreased [LDH (U/L): 896.3±161.5 vs. 1568.3±243.5, CK-MB (U/L): 304.3±72.9 vs. 583.6±81.6], the expressions of p-Akt and SR-B1 were significantly increased (p-Akt/t-Akt: 0.84±0.13 vs. 0.18±0.06, SR-B1/β-actin: 1.23±0.19 vs. 0.09±0.02), with statistically significant differences (allP < 0.05). Compared with OGD+HDL group, necrotic cells in LY294002 group were increased, cell activity was decreased, LDH and CK-MB contents in supernatant were increased, p-Akt and SR-B1 expressions were decreased; there was no statistical difference between LY294002 group and OGD group. There was no significant difference in cell apoptosis among the 4 groups.Conclusions HDL has protective effect on the mice myocardial cells. The mechanism may be related with the up regulation of the expression of SR-B1 protein by the activation of PI3K/Akt pathway.

Expression of scavenger receptor class type B1 in esophageal squamous cell carcinoma and its effects on cell proliferation and invasion / 中华消化杂志

Objective To explore the expression of scavenger receptor class type B 1 (SR-B1) in esophageal squamous cell carcinoma (ESCC) ,and its effects on proliferation and invasion of ESCC cells . Methods From May 2012 to August 2017 ,63 ESCC patients who underwent surgical resection in Henan Provincial People′s Hospital were enrolled and ESCC tissues and corresponding normal tissues were collected . The expression of SR-B1 protein in collected tissues and ESCC cells were detected by immunohistochemistry and Western blotting .ESCC EC1 cells and TE1 cells were transfected with SR-B1 small interfering RNA (siRNA) ,control siRNA ,pcDNA3 .1 and pcDNA3 .1-SR-B1 ,respectively .After transfection ,the expression of SR-B1 protein was examined by Western blotting .The cell proliferation , cell cycle and invasion ability of ESCC EC1 cells and TE1 cells after transfection were determined by cell counting kit-8 (CCK-8) assay ,flow cytometry and Transwell chamber .Chi-square test and t test were performed for statistical analysis .Results The positive rate of SR-B1 protein expression in ESCC tissues was 60 .3％ (38/63) ,which was higher than that of corresponding normal tissues (30 .2％ ,19/63) ,and the difference was statistically significant (χ2=11 .565 , P=0 .001) .The expression of SR-B1 protein in ESCC cells Eca109 ,EC9706 ,EC1 ,TE1 and KYSE70 were 0 .244 ± 0 .012 ,0 .285 ± 0 .018 ,0 .455 ± 0 .016 ,0 .479 ± 0 .019 and 0 .390 ± 0 .022 ,respectively ,which were all higher than that of normal esophageal epithelial cell Het-1A (0 .027 ± 0 .011) ,and the differences were statistically significant (t=23 .252 ,21 .633 ,39 .081 ,36 .010 and 25 .591 ;all P<0 .01) .The expression of SR-B1 protein in ESCC EC1 and TE1 was downregulated by SR-B1 siRNA . The downregulation of SR-B1 protein expression suppressed the proliferation of ESCC EC1 cells and TE1 cells ,increased the percentage of cells at G0/G1 phase of ESCC EC1 cells and TE1 cells ((64 .92 ± 1 .68)％ and (64 .34 ± 0 .94)％ ) ,and reduced the invasion abilities of EC1 and TE1 cells (33 .33 ± 7 .51 and 21 .67 ± 4 .04) .The upregulation of SR-B1 expression promoted the proliferation of ESCC EC1 cells and TE1 cells ,reduced the percentage of cells at G0/G1 phase ((31 .72 ± 1 .30)％ and (33 .12 ± 1 .04)％ ) ,and enhanced cell invasion abilities (285 .33 ± 28 .10 and 247 .33 ± 28 .29) .Conclusions The overexpression of SR-B1 in ESCC may be associated with the occurrence and development of ESCC .Treatment targeting SR-B1 may be a novel therapeutic strategy in ESCC .

Effect of Herbal-cake-separated Moxibustion on Blood Lipid Levels and Expression of Hepatic PPARγ and SR-B 1 Proteins and Genes in Hyperlipidemia Atherosclerosis Rabbits / 针刺研究

OBJECTIVE: To observe the effect of herbal-cake-separated moxibustion on blood lipid levels and expression of peroxisome proliferator-activated receptor γ (PPARγ) and scavenger receptor B 1 (SR-B 1) proteins and genes in liver of hyperlipidemia atherosclerosis rabbits, so as to explore its mechanism underlying anti-atherosclerosis formation. METHODS: Forty male New Zealand white rabbits were randomly divided into normal control, model, moxibustion and Simvastatin groups (n＝10 rabbits in each group). The hyperlipidemia atherosclerosis model was established by high cholesterol diet and propylthiouracil for 12 weeks. Herbal-cake-separated moxibustion was applied to "Juque" (CV 14), and bilateral "Tianshu" (ST 25), "Fenglong" (ST 40) (point group 1), and bilateral "Xinshu" (BL 15), "Ganshu" (BL 18) and "Pishu" (BL 20) (point group 2). The two groups of points were used alternately. Simvastatin (1.96 mg•kg－1•d－1) mixed in the forage was given to rabbits of the Simva-statin group. Both moxibustion and medication treatments were given once daily for continuous 4 weeks. Total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C) and high density lipoprotein cholesterol (HDL-C) levels in plasma were detected by using an automatic biochemistry analyzer. The expression levels of PPARγ and SR-B 1 proteins and genes in the hepatic tissue were determined by Western blot and reverse transcription-polymerase chain reaction, separately. RESULTS: After modeling, plasma TC, TG and LDL-C levels in the model group were significantly increased (P<0.01), and the levels of plasma HDL-C and hepatic PPARγ and SR-B 1 protein and mRNA expression were obviously down-regulated relevant to the normal group (P<0.01). Compared with the model group, plasma TC, TG and LDL-C levels were significantly decreased (P<0.01), and plasma HDL-C and hepatic PPARγ and SR-B 1 protein and mRNA levels were significantly up-regulated in the two treatment groups (P<0.01, P<0.05). CONCLUSION: Herbal-cake-separated moxibustion can regulate blood lipid levels and suppress hyperlipidemia-induced decrease of expression of hepatic PPARγ and SR-B 1 proteins and genes in hyperlipidemia atherosclerosis rabbits, which maybe contribute to its action in anti-atherosclerosis through promoting reversal of cholesterol.

Effect of Electroacupuncture on Expression of Scavenger Receptor A I in Peritoneal Macrophages of Atherosclerotic Rabbits / 针刺研究

OBJECTIVE: To observe the effect of electroacupuncture (EA) on the expression of scavenger receptor A I (SR-A I) protein and mRNA in peritoneal macrophages of atherosclerosis (AS) rabbits. METHODS: A total of 26 New Zealand rabbits were used in the present study, among them, 7 were randomly selected to constitute a blank control group, and the rest 19 rabbits were fed with high-fat diet combined with carotid artery balloon injury for establishing AS model. Eight weeks after high-fat forage feeding, one high-fat fed rabbit and one normal rabbit were randomly selected for verifying the success of modeling. HE staining was employed to examine pathological changes of the common carotid artery after paraffin section. Then the rest 18 rabbits were randomly divided into model, EA and medication groups (n=6 rabbits in each). EA (2 Hz, 1 mA) was applied at "Guanyuan"(ST 25)and bilateral "Zusanli"(ST 36) and "Neiguan" (PC 6) for 20 min. The rabbits of the medication group were treated by gavage of Atorvastatin Calcium (1 mg•kg-1• d-1), and those of the model group treated by gavage of distilled water (2 mL•kg-1•d-1). The treatment was given once daily for 4 weeks, with one day's interval between every two weeks. Three days before termination of the experiments, sterile starch solution (3-4 mL/animal) was injected into the peritoneal cavity, and at the end of the experiment, the peritoneal fluid was collected and centrifugated to be cultivated in Dulbecco's Modified Eagle Medium (DMEM) for collecting the macrophages. The expression of SR-A I protein and mRNA in macrophages was assayed by Western blot and PCR, respectively. RESULTS: HE staining displayed an injury of the inner membrane of the carotid artery marked by observable atherosclerotic plaques, interruption, thickening and uplift, inflammatory cell infiltration, etc. in the AS model rabbits which were relatively milder in both EA and medication groups. The expression levels of SR-A I protein and mRNA were significantly up-regulated in the model group relevant to the blank control group (P0.05). CONCLUSION: EA intervention may improve the severity of atherosclerosis to a certain degree in AS rabbits, which is possibly associated with its effect in inhibiting the expression of SR-A I protein and mRNA in the peritoneal macrophages.

Gene expression of liver nuclear receptor LRH-1 and scavenger receptor class B type I gene in cholesterol gallstone mice / 中国现代普通外科进展

Objective:To investigate the expressions of liver receptor homolog 1 (LRH-1) and scavenger receptor B type Ⅰ (SRBI) gene in cholesterol gallstone (CGS) mice.Methods:Forty C57BL/6 mice,20 with cholesterol gallstone (GS)and 20 controls without gallstones (GSF) were enrolled in this study.mRNA and protein expression of LRH-1 and SRBI genes were determined by RT-PCR and Western blot.Results:Gallbladder walls of GS mice were thicker with increased stromal granulocyte infiltration.The expression levels of LRH-1 genes were significantly higher in GS mice than in controls(P＜0.01).The expression levels of SRBI genes were also significantly higher in GS mice than in controls(P＜0.01).Conclusion:The increased expression of LRH-1 and SRBI gene may be related to GS disease.

Gene expression of liver nuclear receptor LRH-1 and scavenger receptor class B type I gene in cholesterol gallstone mice / 中国现代普通外科进展

Objective:To investigate the expressions of liver receptor homolog 1 (LRH-1) and scavenger receptor B type Ⅰ (SRBI) gene in cholesterol gallstone (CGS) mice.Methods:Forty C57BL/6 mice,20 with cholesterol gallstone (GS)and 20 controls without gallstones (GSF) were enrolled in this study.mRNA and protein expression of LRH-1 and SRBI genes were determined by RT-PCR and Western blot.Results:Gallbladder walls of GS mice were thicker with increased stromal granulocyte infiltration.The expression levels of LRH-1 genes were significantly higher in GS mice than in controls(P＜0.01).The expression levels of SRBI genes were also significantly higher in GS mice than in controls(P＜0.01).Conclusion:The increased expression of LRH-1 and SRBI gene may be related to GS disease.

Study on SCARB1 mediated coagulation dysregulation in recipients after liver xenotransplantation / 器官移植

To investigate the changes in the expression levels of scavenger receptor class B member 1 (SCARB 1) in the liver tissues before and after liver xenotransplantation and analyze the relationship between the variations in the SCARB1 expression and coagulation regulating dysfunction in the recipients.Methods The Wuzhishan miniature pig with α-1,3-galactosyltransferase gene-knockout(GTKO) was utilized as the donor and Macaca thibetana was chosen as the recipient.Heterotopic auxiliary liver xenotransplantation models were established.The liver tissue specimen was collected before and after liver xenotransplantation.Primary hepatocytes were extracted from the pig using collagenase digestion method.Human peripheral blood mononuclear cells were obtained by immunomagnetic bead sorting.These two types of cells were co-cultured and supplemented with human plasma to establish cell models with coagulation regulating dysfunction following liver xenotransplantation.Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) and Western blot were performed to quantitatively measure and statistically compared the expression levels of messenger ribonucleic acid (mRNA) and protein of SCARB1 in the tissue and cell samples.At the cellular level,the expression of SCARB 1 was interfered by lentiviral vector.The coagulation time was detected to validate the effect upon coagulation function.Results The expression levels of SCARB1 mRNA and protein were significantly down-regulated after liver xenotransplantation (both P＜0.05).In the cell models,the expression levels of SCARB1 mRNA and protein in the porcine hepatocytes co-cultured with human monocytes were significantly down-regulated compared with those in porcine hepatocytes without intervention (both P＜0.05).Compared with the non-intervention group,the coagulation time was significantly prolonged after the expression of SCARB1 was interfered by lentiviral vector (P＜0.05).Conclusions The down-regulated expression of SCARB1 in the liver graft is one of the main causes of mediating coagulation regulating dysfunction.Intervention of SCARB1 expression contributes to resolve the coagulation regulating dysfunction in the recipients after liver xenotransplantation.

The role of SR-BⅡ in the oxidized low density lipid induced foam cell formation promoted by porphyromonas gingivalis / 实用口腔医学杂志

Objective:To investigate the role of scavenger receptor BⅡ(SR-BⅡ) in the oxidized low density lipid(ox-LDL) induced foam cells formation promoted by porphyromonas gingivalis(P.g).Methods:Peritoneal macrophages were stimulated with P.g and ox-LDL,and then foam cells formation were checked.The expression of SR-BⅡ was detected by Western blot and real-time PCR.Next,siRNA targeting SR-BⅡ was used to detect the change of foam cells formation.Results:After being stimulated with P.g and ox-LDL,the foam cells formation was significantly increased.During the process of foam cells formation,P.g infection increased the expression of SR-BⅡ.And the knockdown of SR-BⅡ by siRNA significantly reduced the foam cells formation.Conclusion:P.g infection can increase the expression of SR-BⅡ and the regulation of SR-BⅡ expression can change the foam cells formation.

Platelet Rich Plasma (PRP) Produces an Atherofibrotic Histophenotype During Craniofacial Bone Repair Due to Changes of Immunohistochemical Expression of Erk1/2, p38α /β, Adiponectin and Elevated Presence of Cells Exhibiting B-scavenger Receptor (CD36+)

Abstract The platelet-extracellular matrix interaction in platelet rich plasma (PRP) through thrombospondin receptor-CD36 induces the secretion of growth factors responsible for cellular proliferation and differentiation during the repair process. Since CD36 also acts as a class B-scavenger-receptor for development of foam-like cells and mitogen-activated kinases, such as Erk1/2 and p38α/β, are important proteins activated by platelet growth factor, the aim of this study was to evaluate the immunohistochemical presence of CD36, Erk1/2, p38α/β during the bone repair treated and non-treated with PRP and to compare these results with the histomorphometry of repair. Simultaneously, the immunopresence of adiponectin was analyzed, which may contribute to osteogenesis at the same time it inhibits fibrosis and impairs adipogenesis and foam cell formation in the medullary area. An artificial bone defect measuring 5×1 mm was produced in the calvaria of 56 Wistar rats. The defects were randomly treated with autograft, autograft+PRP, PRP alone and sham. The animals were euthanized at 2 and 6 weeks post-surgery. Data were analyzed by ANOVA followed by non-parametric test Student Newman-Keuls (p<0.05) for histomorphometric and immunohistochemical interpretation. The results revealed that in specimens that received PRP the immunopositivity for Erk1/2, p38α/β and CD36 proteins increased significantly while the immunohistochemical expression of adiponectin decreased simultaneously. There was also an accentuated reduction of bone matrix deposition and increase of the medullary area represented by fibrosis and/or presence of foam-like cells, which exhibited immunophenotype CD36+adiponectin. The findings of this study suggest that PRP acted as an inhibitor of osteogenesis during the craniofacial bone repair and induced a pathological condition that mimics an atherofibrotic condition.

Resumo A interação da matriz extracelular-plaquetas no plasma rico em plaquetas (PRP) através de receptor trombospondina CD36 induz a secreção de fatores de crescimento responsáveis pela proliferação e diferenciação celular durante o processo de reparo. Uma vez que o CD36 também age como receptor scavenger de classe B para o desenvolvimento de células do tipo espuma, e as quinases ativadas por mitógenos, tais como ERK1/2 e p38α/β, são importantes proteínas ativadas por fator de crescimento das plaquetas, o objetivo deste estudo foi avaliar a presença imunoistoquímica de CD36, ERK1/2, p38α/β durante o reparo ósseo tratado e não-tratado com PRP e comparar estes resultados com a histomorfometria do reparo. Simultaneamente, analisou-se a imunopresença da adiponectina, que pode contribuir para osteogênese ao mesmo tempo que inibe a fibrose e prejudica a formação de células tipo espuma/xantomatosas na área medular. Um defeito artificial de osso medindo 5×1 mm foi produzido na calvária de 56 ratos Wistar. Os defeitos foram tratados aleatoriamente com auto-enxerto, enxerto autógeno+PRP, PRP apenas e sham. Os animais foram sacrificados 2 e 6 semanas pós-cirurgia. Os dados foram examinados por meio de ANOVA, seguido pelo teste não-paramétrico Student Newman-Keuls (p<0,05) para a interpretação histomorfométrica e imunoistoquímica. Os resultados revelaram que as amostras que receberam PRP aumentaram significativamente a imunopositividade para as proteínas ERK1/2, p38α/β e CD36, simultaneamente à diminuição de expressão imunoistoquímica da adiponectina. Houve também expressiva redução de deposição de matriz óssea e aumento da área medular representada por fibrose e/ou presença de células do tipo espuma que apresentaram imunofenótipo CD36 + adiponectina. Estes resultados sugerem que o PRP atuou como um inibidor da osteogênese durante o reparo ósseo craniofacial e induziu uma condição patológica que mimetiza uma condição aterofibrótica.

In vitro effect and mechanistic study of compound E23869 on lipid metabolism / 药学学报

This study was designed to identify inducers of ATP-binding cassette transporter A1(ABCA1) and CD36 and lysosomal integral membrane protein-II analogous-1(CLA-1) and to evaluate the in vitro effect of the active compound on lipid metabolism. Among 20000 compounds screened, E23869 was found as a positive hit using cell-based high throughput screening models. The up-regulating activities of E23869 in ABCA1p-LUC and CLA-1p-LUC HepG2 cells were 196% and 198%, respectively. The EC50 values of E23869 in ABCA1p-LUC and CLA-1p-LUC HepG2 cells were 0.25 μmol·L-1 and 0.66 μmol·L-1, respectively. E23869 significantly upregulated the protein levels of ABCA1, scavenger receptor class B type I (SR-BI)/CLA-1 and ATP-binding cassette transporter G1(ABCG1) in both macrophages RAW264.7 and L02 cells by Western blotting analysis. Foam cell assay showed that E23869 inhibited lipids accumulations in macrophages RAW264.7. Cholesterol efflux assay showed that E23869 induced HDL-mediated cholesterol efflux in macrophages RAW264.7. Moreover, E23869 up-regulated ABCA1, SR-BI/CLA-1 and ABCG1 expressions through activation of PPARα and PPARγ. In addition, E23869 weakly promoted in vitro differentiation of mouse preadipocytes 3T3-L1. In conclusion, E23869 up-regulated ABCA1, SR-BI/CLA-1 and ABCG1 expressions to promote cholesterol efflux, which is a good leading compound for regulation of lipid metabolism.

The value of serum soluble cluster differentiation CD163 to the prognosis of patients with stroke associated pneumonia / 中国中西医结合急救杂志

ObjectiveTo estimate the value of serum soluble scavenger receptor CD163 (sCD163) to the prognosis of patients with stroke associated pneumonia (SAP).Methods A prospective study was conducted. The clinically suspected SAP patients admitted to Department of Intensive Care Unit (ICU) and Emergency ICU (EICU) in Zhejiang Province Zhuji City Chinese Medicine Hospital from February 2014 to January 2015 were all enrolled. According to clinical pneumonia severity index (PSI), they were divided into SAP group and non SAP group according to the presence or absence of SAP, the patients of SAP group were subdivided into mild SAP group (PSI grade Ⅰ-Ⅲgrade) and severe SAP groups (PSI grade Ⅳ-Ⅴ grade) and according to the 28-day prognosis, the patients were subdivided into hospitalized death group and survival group. The clinical data were collected, including gender, age, history of stroke presence or absence, present stroke pattern, National Institutes of Health Stroke Scale score (NIHSS) on the day of stroke suspect diagnosis, Glasgow coma scale (GCS) score, acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ) score, clinical pulmonary infection score (CPIS), PSI score, body temperature, result of chest X-ray film, oxygenation index (PaO2/FiO2), result of sputum culture, serum levels of white blood cell (WBC) and C-reactive protein (CRP) in a period of 7 days after the suspected diagnosis, PCT, sCDl63 levels on days 1, 3, 5, 7 days, length of stay in ICU, the total time of hospitalization and 28-day survival situation, etc. The ability of each index to evaluate the prognosis of SAP was analyzed by the receiver operating characteristic curve (ROC curve). The risk factors influencing the prognosis of SAP patients were analyzed by multivariate logistic regression analysis.Results ① Seventy-eight patients were finally enrolled in the study, 44 patients were diagnosed as SAP, 34 were non SAP. In 44 patients with SAP, there were 28 cases of severe SAP and 16 cases of mild SAP. On the first day of the suspected diagnosis, the NIHSS score [13 (7, 22) vs. 8 (4, 17), the CPIS score [6 (4, 9) vs. 4 (3, 5), sCD163 [mg/L: 0.80 (0.59, 1.32) vs. 0.33 (0.22, 0.46)], CRP [mg/L: 84.2 (50.8, 114.9) vs. 51.4 (26.2, 79.9)] and 28-day mortality [38.6% (17/44) vs. 11.8% (4/34)] in SAP group were significantly higher than those in non SAP group (allP 0.05). ② The levels of sCD163 reached the peak value on the third day after the suspected diagnosis among SAP group and non SAP group, mild SAP group and severe SAP group, survival group and death group and then began to fall; the levels of sCD163, WBC, CRP, PCT within 7 days in SAP, severe SAP and death groups were higher than those in non SAP, mild SAP and survival groups (allP < 0.05). ③ROC curve analysis indicated: sCDl63 showed a better capacity for evaluating the 28-day prognosis of SAP [ROC curve (AUC) =0.673, 95% confidence interval (95%CI) = 0.515-0.807, sensitivity and specificity were 41.2% and 96.3% respectively and the cut-off was 2.65 mg/L]. However, the levels of other inflammatory indexes and scores on the first day after the suspected diagnosis had no value for early prognosis of SAP. The multiple logistic regression analysis showed that the level of sCD163 on the first day after the suspected diagnosis was the independent risk factor of death in hospital of SAP patients [dominance ratio (OR = 1.27, 95%CI = 1.06-1.52,P < 0.05]. Age (OR = 1.04, 95%CI = 1.01-1.06,P = 0.015), NIHSS score (OR = 2.86, 95%CI = 1.64-4.92,P = 0.010), CPIS score (OR = 1.52,95%CI = 1.28-1.90,P < 0.001) and APACHEⅡ score (OR = 2.06, 95%CI = 1.53-3.07,P < 0.001) were also the risk factors of influencing the death of patients with SAP.Conclusions Early sCD163 level is an independent risk factor in predicting the 28-day mortality of patients with SAP, and it has a certain value for the prognosis of SAP.

Expression and its significance of caveolin-1 in liver and gallbladder of gallstone mice model / 中华消化杂志

Objective To investigate the role and possible mechanism of caveolin-1 (CAV1) in the forming of cholesterol gallstone in mice fed with lithogenic diet.Methods Cholesterol gallstone susceptible C57BL/6 mice were study objects.The mice of control group (n=6) and experiment group (n=6) were fed with normal diet and lithogenic diet for four weeks respectively.The condition of cholesterol gallstone forming,changes of serum lipid and bile composition were measured,and the expressions of CAV1 and scavenger receptor classB member Ⅰ (SR-BⅠ) at mRNA and protein level in the liver and gallbladder were detected by realtime-polymerase chain reaction and Western blot,respectively.The t test was performed for mean comparsion between the two groups.Results The incidence rate of gallstone in experimental group was 100％ after fed with lithogenic diet for four weeks,the lipid level significantly increased,and the proportion of cholesterol in bile raised and bile salt decreased.Compared with those of control group,the expressions of CAV1 at mRNA and protein level in the liver and gallbladder tissues siginificantly decreased (in liver tissue,mRNA 0.53 ± 0.13 vs 1.00 ± 0.32,t =3.330,protein level 0.39 ± 0.07vs 0.92±0.06,t=10.280; in gallbladder tissue,mRNA 0.40±0.22 vs 1.00±0.22,t=3.823,protein level 1.04±0.07 vs 1.34 ± 0.04,t =6.367,all P＜0.01).There was no significant difference in the relative expression of SR-BⅠ at mRNA and protein level in the liver and gallbladder tissues between the mice of experiment group and control group.Conclusion The changes of CAV 1 expression at mRNA and protein level in liver and gallbladder tissues may affect lipids metabolism and cholesterol transportation in liver and gallbladder tissues of experiment mice,which might play an important role in the formation of cholesterol gallstone.